Carolina Rinse Solution Research Articles

Brief topical and intraluminal use of Carolina rinse solution does not attenuate experimental ischemia and reperfusion injury in rabbit jejunum

ABSTRACT Fifteen New Zealand adult rabbits were randomly allocated into three groups: Sham-operated (group A), Ischemia and Reperfusion (group B) and Carolina Rinse Solution (CRS) (group C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes before reperfusion, the bowel lumen was filled with CRS, and the segment immersed in CRS. Necrosis and loss of integrity of the villi were visible in groups B and C. Edema of the submucosa and circular muscle was observed in all groups. Hemorrhage was observed in different layers for groups B and C, but group C showed more severe hemorrhage in different layers during reperfusion. All groups showed polymorphonuclear leukocyte infiltration on the base of the mucosa, submucosa, and longitudinal muscle, in addition to polymorphonuclear leukocytes margination in the mucosal and submucosal vessels. Necrosis of enterocytes, muscles, crypts of Lieberkühn and myenteric plexus was observed in groups B and C during reperfusion. Topical and intraluminal Carolina Rinse Solution did not attenuate the effects of ischemia and reperfusion in the small intestine of rabbits.

Topical and intraluminal Carolina Rinse Solution in p44/42 and p38 MAP Kinase activation profile in rabbit jejunum after ischemia and reperfusion

The effects of topical and intraluminal Carolina Rinse Solution (CRS) in p44/42 (ERK 1/2) and p38 MAPK activation profile, in rabbit jejunum after ischemia and reperfusion (I/R), were investigated in this study. Fifteen New Zealand rabbits were allocated in three groups: Sham-operated (A), Ischemia and reperfusion (B) and CRS (C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes prior to reperfusion, the bowel lumen was filled with CRS, and the segment was immersed in CRS until reperfusion onset. Ischemia and reperfusion stimulated the phosphorylation of the p44/42 MAPK and p38 MAPK pathways in some layers of jejunum. Progressive activation of p44/42 MAPK was chiefly localized in the crypts of Lieberkühn, circular and longitudinal muscle layers, whereas p38 MAPK was prominently activated in myenteric plexus and both muscle layers. The results of this research indicate that the chosen model of topical and intraluminal CRS does not interfere in p44/42 and p38 MAPK activation profile in rabbit jejunum subjected to I/R.

Effects of Carolina rinse solution, dimethyl sulfoxide, and the 21-aminosteroid, U-74389G, on microvascular permeability and morphology of the equine jejunum after low-flow ischemia and reperfusion

To evaluate effects of Carolina rinse solution, dimethyl sulfoxide (DMSO), and 21-aminosteroid, U-74389G, on microvascular permeability and morphology of the equine jejunum after low-flow ischemia and reperfusion. 20 healthy adult horses. Under anesthesia, full-thickness biopsy specimens of a distal portion of the jejunum were obtained for baseline measurements. In addition to a control segment, 2 jejunal segments were identified as sham-operated or experimental segments. Experimental segments underwent 60 minutes of low-flow ischemia and 3.5 hours of reperfusion. Treatments were as follows: U-74389G (3 mg/kg, IV; 6 horses), DMSO (20 mg/kg, IV; 6) diluted in 1 L of saline (0.9% NaCl) solution, local perfusion (via jejunal artery) of Carolina rinse solution (0.5 mL/kg; 4), and local perfusion of lactated Ringer's solution (0.5 mL/kg; 4). Jejunal microvascular permeability was significantly lower after treatment with Carolina rinse solution or DMSO, compared with U-74389G or lactated Ringer's solution treatments. After DMSO treatment, serosal- and submucosal-layer edema was significantly increased in experimental segments, compared with control or sham-operated segments; however, edema increases were significantly less than for lactated Ringer's solution or U-74389G treatments. Significant decreases in intestinal wet weight-to-dry weight ratio were found following Carolina rinse solution or DMSO treatments, compared with lactated Ringer's solution or U-74389G treatments. Edema formation and leukocyte infiltration in jejunal segments of horses treated with lactated Ringer's solution or U-74389G were increased, compared with Carolina rinse solution or DMSO treatments. Carolina rinse solution and DMSO may be protective against ischemia-reperfusion injury in the equine jejunum.

Role of free radicals in failure of fatty liver grafts caused by ethanol

Alcohol is associated with accidental deaths and suicides leading to organ donation, and hepatic steatosis is an important risk factor for initial poor function and failure of human liver grafts. Mechanisms of fatty graft failure are not fully understood, but increased oxidative stress may be a major factor. To characterize the role of free radical stress and the efficacy of antioxidant treatments in fatty liver graft injury, donors for orthotopic rat liver transplantation were treated chronically (3 or more weeks) and acutely (single dose) with ethanol. After transplantation, necrosis and alanine aminotransferase release were threefold to fourfold higher in recipients of fatty grafts from donors treated with ethanol either acutely or chronically compared with findings for recipients of grafts from untreated donors. Moreover, graft survival decreased from nearly 100% to less than 20%. Free radical adducts, as measured by electron spin resonance spectroscopy, were detected in the blood and bile of rats receiving fatty grafts caused by ethanol. Markers of lipid peroxidation also increased after transplantation. Destruction of Kupffer cells with gadolinium chloride decreased free radical production and improved graft survival. Leukocyte adhesion increased beginning early after implantation, and adherent white blood cells obtained from transplanted fatty livers produced the same free radical species as were detected in blood. Therefore, Kupffer cells and adherent white blood cells are important sources of free radicals. Free radicals not only damage fatty grafts directly but also lead to enhanced inflammation and disturbed microcirculation. Delivery of superoxide dismutase-1 and superoxide dismutase-2 genes, free radical–scavenging polyphenols, and antioxidant-containing Carolina Rinse solution reduced injury and improved survival of fatty grafts caused by ethanol. Taken together, these findings indicate that free radicals increase in fatty grafts after transplantation and play an important role in injury of fatty grafts obtained from ethanol-exposed donors. Treatment of fatty donor livers with antioxidants and free radical scavengers may thus be an effective clinical therapy to prevent failure of fatty grafts.

Evaluation of Carolina Rinse solution as a treatment for ischaemia reperfusion of the equine jejunum.

Ileus and peritoneal adhesions are the most common complications following surgery for small intestinal obstruction. Carolina Rinse (CR) has been shown to decrease reperfusion injury in intestine and other organs. CR decreases intestinal inflammation and subsequent scarring associated with reperfusion injury. CR was infused intra-arterially and applied topically just prior to reperfusion in jejunum exposed to experimental ischemia. Vascular permeability, neutrophil accumulation and serosal scarring were compared in treated and untreated intestine. CR maintained a normal osmotic reflection coefficient and decreased migration of neutrophils into the serosa during reperfusion. After 10 days, treated intestine was normal in appearance with a trend toward less serosal scarring and fibroblast proliferation. There was a significant decrease in fibroplasia at biopsy sites in treated intestine. Arterial perfusion combined with topical application of CR during jejunal ischaemia decreases immediate reperfusion injury and limits post operative scarring. CR should be used as a local perfusate rather than a systemic treatment; it may best be applied topically and intraluminally to avoid damaging mesenteric arteries. CR should be considered an adjunct treatment as part of overall surgical management and post operative care.

Detrimental effects of controlled reperfusion on renal function after porcine autotransplantation are fully compensated by the use of Carolina rinse solution

Detrimental effects of controlled reperfusion on renal function after porcine autotransplantation are fully compensated by the use of Carolina rinse solution

Detrimental effects of controlled reperfusion on renal function after porcine autotransplantation are fully compensated by the use of Carolina rinse solution.

Despite extensive efforts in the fields of donor selection and management, standardisation of organ retrieval procedures, storage solutions, and novel immunosuppressive protocols, the rates of delayed graft function (DGF) after renal transplantation have been stagnating between 30% and 50%. As DGF exerts negative influences on acute rejection episodes and long-term organ function, the early phase of transplantation immediately following reperfusion deserves special interest. Several studies on machine-controlled reperfusion showed promising results in various organs, in experimental and clinical settings. Moreover, the flushing of organs with Carolina rinse solution (CR) immediately prior to reperfusion has been proven beneficial and is being clinically applied in human liver transplantation in recognised departments. In our study, we set up an autogenic porcine kidney transplantation model and assessed the normal values (control group) for creatinine clearance (ClCr) and urine output per hour (U/h) after "standard" reperfusion similar to clinical transplantation. Subsequently, kidneys of the experimental group 1 were reperfused at a blood pressure (RR) under the systemic level by means of a roller pump. Group 2 kidneys were rinsed with CR before controlled reperfusion, analogous to group 1. Both groups were compared with each other and with the assessed normal values. Our findings for Group 1 are that pressure-reduced reperfusion negatively affected immediate graft function. ClCr was reduced from 9.9 (control group) to 3.4 ml/min, U/h from 233 to 132 ml ( P<0.05). Group 2 showed that rinsing the kidneys with CR before reperfusion improved functional parameters highly significantly, compared with group 1 (ClCr: 13.5 vs 3.4 ml/min, U/h: 384 vs 132 ml; P<0.05) and even showed a positive trend compared with the control group (ClCr: 13.5 vs 9.9 ml/min, U/h: 384 vs 233 ml; P=0.0546). We can conclude that in a model of porcine renal autotransplantation, pressure-reduced reperfusion via a roller pump is detrimental to early kidney graft function. The flushing of organs with CR prior to controlled reperfusion significantly improves ClCr as well as urine output.

Carolina rinse solution minimizes kidney injury and improves graft function and survival after prolonged cold ischemia.

Kidney damage caused by cold ischemia-reperfusion injury promotes adverse outcomes after renal transplantation. The purpose of this study was to determine whether Carolina rinse solution (CRS) used at the end of cold ischemic storage decreases kidney injury and improves graft function and survival. Inbred male Lewis rats were used as donors and recipients. Left kidneys were removed from donor rats, infused with cold University of Wisconsin solution, and stored for 24, 30, or 48 hr at 0-1 degrees C. Just before implantation, kidneys were flushed with either Ringer's solution or CRS at 35-37 degrees C or were not treated. Kidneys were then transplanted into recipient rats with removal of both native kidneys. Survival and renal function were analyzed over a 14-day postoperative period. Among rats receiving kidneys after 24-hr cold storage, creatinine clearance was 75% greater in rats transplanted with kidneys flushed with CRS compared with Ringer's solution or nontreatment. In animals receiving kidneys after 30-hr cold storage, recipient survival after CRS was significantly higher than with Ringer's solution or without flushing (80% vs. 25% and 17%, respectively). However, CRS failed to prevent renal graft failure after 48 hr of cold storage (14% survival with CRS vs. 0% with Ringer's solution). In separate ex vivo studies, nonviable cell nuclei were labeled by trypan blue after cold preservation and brief warm reperfusion. CRS decreased podocyte and peritubular endothelial cell killing associated with cold ischemia-reperfusion injury. Flushing renal explants with warm CRS before implantation diminishes cold ischemia-reperfusion injury and improves the function and survival of transplanted kidneys.

Evaluation of parenchymal and nonparenchymal cell injury after different conditions of storage and reperfusion.

We used the isolated perfused rat liver model (IPRL) to assess parenchymal and nonparenchymal cell integrity after different conditions of storage and reperfusion. Two studies were performed. In study 1, the IPRL was applied to evaluate the effects of 30 min of normothermic reperfusion with Elohes solution, enriched William's medium (Wif), or Carolina rinse solution (CRS) following 24 h of cold preservation in high-K+ or high-Na+ UW solution. As indicated by creatine kinase-BB (CK-BB) release, reperfusion with CRS provided greater protection of endothelial cells after storage in high-K+ UW solution than after storage in high-Na+ UW solution. In study 2, livers were cold-preserved (24 h, 4 degrees C) in either high-K+ or high-Na+ UW solution, then flushed with either CRS or Wif solution at room temperature before reperfusion (120 min, 37 degrees C) with 5% albumin-William's medium E. There was no statistical difference between the rinse solutions for bile flow and transaminases release. However, CRS improved bile indocyanine green excretion, which is known to be a marker of parenchymal and nonparenchymal cell integrity. Therefore, we can assume that this rinse solution protects rat liver grafts from reperfusion-induced microvascular damage.

Reperfusion injury after liver preservation for transplantation.

Preservation injury remains an obstacle to greater utilization of liver transplantation therapy. Livers can be preserved a maximum of 24 h in University of Wisconsin solution. After longer times, reperfusion precipitates endothelial cell killing and activation of Kupffer cells (liver macrophages). Together, Kupffer cell activation and endothelial cell killing cause microcirculatory disturbances, leukocyte and platelet adhesion, and a systemic inflammatory response after graft implantation. Down-regulation of Kupffer cells with calcium blockers or pentoxifylline improves graft survival, whereas priming with lipopolysaccharide or alcohol worsens survival. Flushing grafts after storage with Carolina rinse solution containing antioxidants, adenosine, calcium blocker, energy substrates, and glycine at pH 6.5 decreases endothelial cell killing, reduces Kupffer cell activation, and improves graft survival. Understanding of the roles of different cells in storage/reperfusion injury forms the basis for strategies to prolong organ storage, improve graft function, and reduce failure of fatty grafts from alcoholic donors.

Studies of rat lung viability and adenine nucleotide metabolism after death

Prior studies from our laboratory have supported the use of cadaveric lungs for transplantation. In this study we investigated different preservation strategies for lungs retrieved from cadavers 4 hours after circulatory arrest. Seventy-two Sprague-Dawley rats were sacrificed and then ventilated with 100% oxygen for 4 hours. The lungs were then flushed with modified Euro-Collins, University of Wisconsin, or Carolina rinse solution, either alone, with prostaglandin E1, or with prostaglandin E1 plus the free radical scavenger dimethylthiourea. After an additional 4-hour cold storage, the left lung was flushed with trypan blue solution to quantify cell viability, whereas the right lung was used to determine wet-to-dry weight ratios and to measure the levels of the adenine nucleotides adenosine triphosphate, adenosine diphosphate, and adenosine monophosphate by high-performance liquid chromatography. Viability was consistently better in the lungs flushed with Carolina rinse solution; these differences were statistically significant compared with those in the corresponding modified Euro-Collins subgroups (p < 0.005). The addition of prostaglandin E1 to all three preservation solutions improved the total adenine nucleotide levels; this increase was statistically significant for the modified Euro-Collins subgroup (p < 0.005). The total adenine nucleotide levels for the University of Wisconsin subgroups were higher than those for the corresponding modified Euro-Collins subgroups. The highest total adenine nucleotide levels were obtained in lungs flushed with Carolina rinse plus prostaglandin E1. Wet-to-dry weight ratios were always significantly lower in the lungs preserved with University of Wisconsin solution (p < 0.05), with a value similar to that of fresh tissue. The characteristics of the solution used to flush and to store rat cadaveric lungs have an impact on lung viability and adenine nucleotide metabolism. The ideal preservation strategy may allow for lung retrieval from cadavers for safe transplantation.

Amelioration of pulmonary allograft injury by administering a second rinse solution

Objective: The use of rinse solutions before reperfusing liver allografts has been shown to reduce cell death in rats. Carolina rinse solution (an extracellular solution that contains antioxidants, vasodilators, and other substrates that help prevent ischemia-reperfusion injury) has also been shown to improve liver function clinically in liver transplant recipients. This pilot study evaluates the value of a second pulmonary artery flush before reperfusion of a lung graft. Methods: Six groups of Sprague-Dawley rats ( n = 6 each) were subjected to the following: Group 1 lungs were preserved with modified Euro-Collins solution followed by 24 hours of cold ischemia. Group 2 lungs were treated the same as group 1 but reperfused with blood. Group 3 lungs were preserved in Carolina rinse solution followed by 24 hours of cold ischemia. Group 4 lungs were treated the same as group 3 lungs and then reperfused with blood. Lungs in groups 5 and 6 were preserved with Euro-Collins solution, stored cold for 24 hours, and then rinsed with Euro-Collins or Carolina rinse solution, respectively, before reperfusion with blood. Lungs were subsequently stained with trypan blue solution for 5 minutes. Lung blocks were fixed and embedded in water-soluble methacrylate. Trypan blue–stained nuclei in nonviable endothelial cells and alveolar pneumocytes were counted in 10 different fields. Results: Groups 1 and 3, preserved with Euro-Collins and Carolina rinse solutions for 24 hours but not reperfused with blood, had significantly more viable endothelial cells (groups 1 and 3 vs group 2, p < 0.0001; group 3 vs group 4, p < 0.02) and pneumocytes (group 1 vs groups 2 and 4, group 3 versus group 2, p < 0.0001; group 3 vs group 4; p < 0.035) than groups 2 and 4, which were subsequently reperfused with blood. Groups 5 and 6, which received a second rinse, also had significantly more viable endothelial cells ( p < 0.0005) and pneumocytes ( p < 0.0001) than control groups, which were not rinsed before reperfusion. Conclusions: We conclude that damage to pulmonary allografts resulting from prolonged ischemia is accentuated by reperfusion with blood. We also conclude that preservation with a single flush of Euro-Collins or Carolina rinse solution does not offer adequate protection, whereas a second rinse before reperfusion significantly decreases the number of damaged cells within the allograft. (J T HORAC C ARDIOVASC S URG 1996;112:1010-6)

Dual role of Kupffer cell activation and endothelial cell damage in reperfusion injury to livers stored for transplantation surgery.

In rat models of liver preservation, the primary event leading to liver graft failure after cold storage is a reperfusion injury causing damage to sinusoidal endothelial cells and activation of Kupffer cells (KC). After storage for longer than 16 h in University of Wisconsin solution, reperfusion induces rapid endothelial cell killing. Kupffer cell activation also occurs as indicated by cell surface ruffling, degranulation, release of hydrolytic enzymes, generation of oxygen radicals, and increased phagocytosis. Down-regulation of KC activity with nisoldipine or pentoxifylline improves graft survival. Moreover, pretreatment of donors with small amounts of endotoxin to activate KC causes a drastic reduction of graft survival. Together, KC activation and endothelial damage cause marked microcirculatory disturbances after transplantation characterized by reduced and uneven blood flow and increased leucocyte and platelet adhesion. Such events culminate in inflammation, necrosis and fulminant graft failure. Modification of reperfusion conditions can reduce the extent of injury. In particular, flushing livers with Carolina rinse solution (CRS) at the end of storage reduces endothelial cell killing, suppresses KC activation, improves the microcirculation, and increases graft survival. Active ingredients in CRS include antioxidants (allopurinol, desferrioxamine and glutathione), adenosine and slightly acidic pH (6.5). Other potentially important ingredients are nicardipine, a calcium channel blocker, and fructose, glucose and insulin to promote glycolysis. The cytoprotective amino acid, glycine, further improves the performance of Carolina rinse solution. Reperfusion-induced changes to nonparenchymal cells play an essential role in damage to livers preserved for transplantation surgery. Understanding the role of sinusoidal endothelial cells and KC in this injury has led to promising new strategies to prolong organ storage and reduce graft failure.

Effectiveness of Carolina rinse solution after cold ischemic storage of rat livers: biochemical and histological analysis using perfusion model.

Using a rat liver perfusion model, the effectiveness of Carolina rinse solution was assessed for the prevention of reperfusion injury after 48 h of cold storage in UW solution. Transaminase levels (GOT, GPT and LDH) of the perfusate were significantly higher in the Ringer group (17 +/- 8, 17 +/- 9 and 191 +/- 97 IU/l, respectively) than in the Carolina group (6 +/- 4, 5 +/- 4 and 21 +/- 20 IU/l) (p < 0.05). The levels of oxygen consumption were also higher in the Carolina group (233 +/- 54 mm Hg) than in the Ringer group (164 +/- 58 mm Hg) (p < 0.05). Histological examination showed severe parenchymal cell damage in the Ringer group, whereas the damage was slight in the Carolina group. Two newly developed monoclonal antibodies, REC16-11 and REC4-1, which specifically react with rat endothelial cells, were used for immunohistochemical studies of the livers. The endothelial cells of central vein and sinusoids were more severely damaged in the Ringer group than in the Carolina group. The present study suggests that Carolina rinse solution is useful for prevention of liver damage from reperfusion injury after cold storage of the graft for organ transplantation.

The validity of the MEGX test in correlation with histology after orthotopic rat liver transplantation.

Lidocaine metabolism (MEGX test) as an indicator for liver function in the assessment of different degrees of liver disease and as a predictor for liver outcome after transplantation is well established. Since reduced liver function is associated with an alteration in parenchymal and non-parenchymal cells, we evaluated whether MEGX values correlate with histology in an in vivo model of orthotopic rat liver transplantation (ORLT) to assess histological damage without taking biopsy specimens. Livers from syngeneic Lewis rats were transplanted with rearterialization after 15-30 h of cold storage in UW solution and rinsing with Carolina Rinse Solution prior to implantation. Forty-eight hours after transplantation, the MEGX test was performed and metabolites were measured with a commercial kit as described elsewhere. Biopsy specimens were taken and graded three degrees of damage (mild, moderate, and severe) in a double blind fashion by a pathologist. MEGX values were assigned to the histological results. Statistical analyses were done with a Mann-Whitney test (n = 58) for mean values. The mean MEGX values attributed to histologies with a mild, moderate, severe degree of damage were 159.96, 78.46 and 44.42 ng/ml, respectively. When the histological groups were compared with the mean MEGX values, mild vs moderate, mild vs severe and moderate vs severe were significant (P - 0.0001). In conclusion, MEGX values correlate significantly with histological grading in a linear fashion after ORLT. The MEGX test may be of clinical value because it reflects the histological pattern of livers and may reduce the necessity to take biopsy specimens before and after transplantation.

Suppression of lipopolysaccharide-stimulated release of tumor necrosis factor by adenosine: Evidence for A2 receptors on rat kupffer cells†

In liver grafts that will fail as a result of storage injury, reperfusion activates Kupffer cells. Overproduction of tumor necrosis factor by activated Kupffer cells may cause primary graft nonfunction, multiple organ failure and, eventually, death of graft recipients. Carolina rinse solution, adenosine, nisoldipine, pentoxifylline and prostaglandin E1 reduce graft failure from storage/reperfusion injury. To test the hypothesis that these agents act by suppressing cytokine release by activated Kupffer cells, we assessed the effect of each drug on tumor necrosis factor released from cultured rat Kupffer cells stimulated with lipopolysaccharide. Adenosine, nisoldipine and prostaglandin E1 each suppressed lipopolysaccharide-stimulated tumor necrosis factor release. The adenosine A2 receptor agonists. 5-n-ethylcarboxamidadenosine, 2-chloro-adenosine and R-phenylisopropyl adenosine also blocked tumor necrosis factor release in a potency suggestive of A2 receptor activity. Xanthine amine congener, a specific A1 receptor antagonist, failed to reverse the suppression by adenosine of tumor necrosis factor release, whereas CGS15943A, an A2 receptor antagonist, did reverse suppression by adenosine and 5-n-ethylcarboxamidadenosine. CGS15943A had no effect on suppression of lipopolysaccharide-stimulated tumor necrosis factor release by nisoldipine or prostaglandin E1. Dibutyryl-cyclicAMP also suppressed tumor necrosis factor release. Adenosine, 5-n-ethylcarboxamidadenosine, prostaglandin E1 and pentoxifylline increased cyclicAMP levels in cultured Kupffer cells, but nisoldipine did not.(ABSTRACT TRUNCATED AT 250 WORDS)

Glycine improves the in vitro and in vivo efficacy of carolina rinse solution in rat liver transplantation

Glycine improves the in vitro and in vivo efficacy of carolina rinse solution in rat liver transplantation

Protective effects of Carolina rinse solution against reperfusion injury in canine renal autotransplantation.

To assess the effect of Carolina rinse solution on the renal graft function after cold ischemic storage in the University of Wisconsin solution for 48 h, canine renal transplantation was performed. Survival rate was higher in the kidneys rinsed with Carolina rinse solution (4 of 5, 80%) than that of kidneys rinsed with lactate Ringer's solution (0 of 5, 0%). Postoperative serum creatinine level on day 2 was significantly lower in the Carolina group (3.48 +/- 0.47 mg/dl) than that in the Ringer group (5.60 +/- 0.68 mg/dl) (p < 0.01). All of the grafts in the Carolina group functioned immediately. Whereas, in the Ringer group, none of the grafts functioned postoperatively. Histologically, severe renal damage was observed in the kidneys perfused by lactate Ringer's solution. On the other hand, the damage to the grafts was minimal in the Carolina group. With immunohistochemical staining using factor-VIII-related antigen, the glomeruli were more severely damaged in the kidneys perfused by lactate Ringer's solution than by Carolina rinse solution. The present study showed that Carolina rinse solution was effective in the prevention of the kidney graft damage from reperfusion injury.

Warm Carolina rinse solution prevents graft failure from storage injury after orthotopic rat liver transplantation with arterialization

An injury to nonparenchymal cells, characterized by loss of viability of sinusoidal endothelial cells and activation of Kupffer cells, occurs after reperfusion of livers stored for transplantation. Recently, a new solution, Carolina rinse solution, was shown to prevent reperfusion injury to endothelial cells in vitro almost completely and to improve graft survival after orthotopic rat liver transplantation (ORLT) without arterialization. ORLT with arterialization permits longer cold storage of donor livers and more closely models human surgery. Therefore, we evaluated the effects of Carolina rinse solution on graft survival after ORLT with arterialization in syngeneic Lewis rats. Just prior to implantation, donor livers stored in University of Wisconsin (UW) solution were rinsed with 30 ml of Ringer's solution, saline, or Carolina rinse solution at 1 degree-4 degrees C. In livers stored for 15 h and rinsed with Ringer's or saline solution, long-term graft survival was only 8%. Using Carolina rinse solution containing 1 mmol and 200 mumol adenosine per liter, graft survival improved to 40% and 80%, respectively. Graft survival did not improve when using Carolina rinse solution with adenosine omitted or Ringer's solution containing 200 mumol adenosine per liter. Livers were also rinsed with Carolina rinse solution containing 200 mumol adenosine per liter at 28 degrees-30 degrees C rather than at 1 degree-4 degrees C. With warm Carolina rinse solution, survival improved further to 100%, 80%, and 50% after 15, 18, and 21 h of storage. After 18 h of storage, light and electron microscopy demonstrated marked denudation of the sinusoidal lining and activation of Kupffer cells in grafts rinsed with Ringer's solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Warm Carolina rinse solution prevents graft failure from storage injury after orthotopic rat liver transplantation with arterialization

Abstract. An injury to nonparenchymal cells, characterized by loss of viability of sinusoidal endothelial cells and activation of Kupffer cells, occurs after reperfusion of livers stored for transplantation. Recently, a new solution, Carolina rinse solution, was shown to prevent reperfusion injury to endothelial cells in vitro almost completely and to improve graft survival after orthotopic rat liver transplantation (ORLT) without arterialization. ORLT with arterialization permits longer cold storage of donor livers and more closely models human surgery. Therefore, we evaluated the effects of Carolina rinse solution on graft survival after ORLT with arterialization in syngeneic Lewis rats. Just prior to implantation, donor livers stored in University of Wisconsin (UW) solution were rinsed with 30 ml of Ringer's solution, saline, or Carolina rinse solution at 1d̀-4d̀C. In livers stored for 15 h and rinsed with Ringer's or saline solution, long-term graft survival was only 8%. Using Carolina rinse solution containing 1 mmol and 200 μmol adenosine per liter, graft survival improved to 40% and 80%, respectively. Graft survival did not improve when using Carolina rinse solution with adenosine omitted or Ringer's solution containing 200 μmol adenosine per liter. Livers were also rinsed with Carolina rinse solution containing 200 μmol adenosine per liter at 28d̀-30d̀C rather than at 1d̀-4d̀C. With warm Carolina rinse solution, survival improved further to 100%, 80%, and 50% after 15, 18, and 21 h of storage. After 18 h of storage, light and electron microscopy demonstrated marked denudation of the sinusoidal lining and activation of Kupffer cells in grafts rinsed with Ringer's solution. Use of Carolina rinse solution greatly improved endothelial structure but did not reduce Kupffer cell activation. In conclusion, these findings show that Carolina rinse solution substantially improves graft survival after ORLT with arterialization. Adenosine and warm temperature are important factors contributing to efficacy. A mechanism of protection appears to be prevention of reperfusion-induced endothelial cell injury.